Oil Palm Transgenic Research: Challenges, Update, and Future Outlook

Rasid, Omar Abdul; Masura, Subhi Siti; Hanin, Ayub Nor; MASLI, DAYANG IZAWATI ABANG; Bahariah, Bohari; Masani, Mat Yunus Abdul; Parveez, Ghulam Kadir Ahmad

doi:10.1007/978-3-030-22549-0_6

Cited by 4 publications

(2 citation statements)

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“…Oil producing woody species such as palm ( Elaeis guineensis ) and Jatropha curcas are promising sources of biodiesel (Carriquiry et al 2011 ). Current efforts are underway to engineer palm trees with enhanced production of oleic acid and with generally reduced saturated fatty acids (Rasid et al 2020 ). In J. curcas , triacylglycerol (TAG) catabolism was suppressed by downregulating a lipase via RNAi, resulting in higher storage lipid accumulation in seeds (Kim et al 2014 ).…”

Section: Goals and Progress Of Metabolic Engineering Of Woody Plantsmentioning

confidence: 99%

Metabolic engineering in woody plants: challenges, advances, and opportunities

2021

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Woody plant species represent an invaluable reserve of biochemical diversity to which metabolic engineering can be applied to satisfy the need for commodity and specialty chemicals, pharmaceuticals, and renewable energy. Woody plants are particularly promising for this application due to their low input needs, high biomass, and immeasurable ecosystem services. However, existing challenges have hindered their widespread adoption in metabolic engineering efforts, such as long generation times, large and highly heterozygous genomes, and difficulties in transformation and regeneration. Recent advances in omics approaches, systems biology modeling, and plant transformation and regeneration methods provide effective approaches in overcoming these outstanding challenges. Promises brought by developments in this space are steadily opening the door to widespread metabolic engineering of woody plants to meet the global need for a wide range of sustainably sourced chemicals and materials.

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Section: Goals and Progress Of Metabolic Engineering Of Woody Plantsmentioning

confidence: 99%

Metabolic engineering in woody plants: challenges, advances, and opportunities

2021

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“…This was done by silencing two key enzymes, oleoyl-CoA desaturase (EgFAD2) and palmitoyl-ACP-thioesterase (EgPAT), that regulate the composition of oil palm fatty acids [ 6 , 7 , 9 ]. However, the research efforts have yet to result in stable lines, and the development of transgenic oil palm is still highly challenging [ 10 ]. In oil palm, EgFAD2 is present in two copies, namely EgFAD2-1 and EgFAD2-2.…”

Section: Introductionmentioning

confidence: 99%

Multiplex CRISPR/Cas9 gene-editing platform in oil palm targeting mutations in EgFAD2 and EgPAT genes

Bahariah

Masani

Fizree

et al. 2023

Journal of Genetic Engineering and Biotechnology

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Background CRISPR/Cas9 is the most powerful and versatile genome-editing tool that permits multiplexed-targeted gene modifications for the genetic enhancement of oil palm. Multiplex genome-editing has recently been developed for modifying multiple loci in a gene or multiple genes in a genome with high precision. This study focuses on the development of high-oleic oil palm, the primary target trait for healthy low-saturated oil. To achieve this, the fatty acid desaturase 2 (FAD2) and palmitoyl-acyl carrier protein thioesterase (PAT) genes, both of which are associated with fatty acid metabolism biosynthesis pathways in oil palm, need to be knocked out. The knockout of FAD2 and PAT leads to an accumulation of oleic acid content in oil palms. Results A total of four single-guide RNAs (sgRNAs) were designed in silico based on the genomic sequences of EgFAD2 and EgPAT. Using robust plant CRISPR/Cas9 vector technology, multiple sgRNA expression cassettes were efficiently constructed into a single-binary CRISPR/Cas9 vector to edit the EgFAD2 and EgPAT genes. Each of the constructed transformation vectors was then delivered into oil palm embryogenic calli using the biolistic, Agrobacterium-mediated, and PEG-mediated protoplast transformation methods. Sequence analysis of PCR products from 15 samples confirmed that mutations were introduced at four target sites of the oil palm EgFAD2 and EgPAT genes. Single- and double-knockout mutants of both genes were generated, with large and small deletions within the targeted regions. Mutations found at EgFAD2 and EgPAT target sites indicate that the Cas9/sgRNA genome-editing system effectively knocked out both genes in oil palm. Conclusion This technology is the first in oil palm to use CRISPR/Cas9 genome-editing to target high-oleic-associated genes. These findings showed that multiplex genome-editing in oil palm could be achieved using multiple sgRNAs. Targeted mutations detected establish that the CRISPR/Cas9 technology offers a great potential for oil palm.

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